This proposal is a part of a long-term project to understand the organization of higher eukaryotic genomes and the molecular biology of functional chromosome components, such as telomeres and centromeres. Telomeres are the specialized structures that form the termini of eukaryotic chromosomes. These structures are essential for the replication and maintenance of the genome. Telomeres have been extensively studied in lower eukaryotes but considerably less in known about higher eukaryotic telomeres. We have recently isolated a higher eukaryotic telomere from the flowering plant Arabidopsis thaliana, providing the opportunity to study the telomeric regions of this organism as a model system. Two aspects of telomere structure and function will be studied. First, the nature and organization of the DNA sequences that lie adjacent to A. thaliana telomeres will be investigated. Second, the mechanism by which a broken chromosome end acquires a new telomere (i.e., telomere healing) will be studied using a chromosomal mutant of A. thaliana which carries a healed telomere. The healed telomere will be isolated and its structure compared to that of the original chromosomal site to determine how the healing event took place. In addition, the healed telomere, which resides proximal to a centromere, will be investigated as a potential tool for studying A. thaliana centromere structure.